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in vitro grown plants w;is ocli~eved with srntic liydropo~iics systc~l~ contai~ii~ig Ar~~on's
nutrie~it solutio~l.
heric,iic irn~i~joj~nriuiio~i. Genetic tri~~isSor~~~;~tion ol' cli~ckpcu ivas achicvcd by
using Ah14 eupla~its as ll~u startlllg plo~~t ii~atrrial and butli biol~st~c illid Agr~~h~icii~riii~)~-
~iied~ated ~iietliods wcrc employed. A vector, ~RTO(I:GUS-III~ nla~i~tn~~led 111 E, cull was
used 111 b~olisric process. 'fh~s vector li:rd irpill pc~ic ;IS ~lic aclcct;~blc ininrkcr and r~itli\
gem nb the rsporrcr. Prcpar;~t~on of thc cxpl,i~ils, niicrocarricr
preparation and
bombard~ncnr of tlie expl:i~~ts was ci~n-~cd out by al;~nd;lrd j~roccd~~rcs. Largc SIX 01' llic
AM3 cxpla~~t was a li~liitatio~~ in 1111s process ns olily fi.w cxplu~ils could be grouped st tile
center of the petri plate. l'ra~lsl'or~iiation by tliis ~lietliod d ~d ~iot result in creatio~~ of any
transgenlcs as some of the selected slioots d~ed at tlie stri~~gerit seluction bkp.
J~grobatie~.i~~~ri-~~lediated transfor~i~otion rcsuited in relatively higli IYeque~icy of
transgenies as far as 111s TO results wcrc considered. Two binary veclors ~ia~iicly pl-IS
723:Ut and pHS 737:SBTl based 111 stra~n C 58, wcrc used for co-cultivarions. Tl~c former
has 81C'ryIAb as agrono~iiic;illy important gcne, ~lptll as thc seiectio~~ 111;lrker and iridA
gclie as ~.cportcr \\l~ere i~s tllc later binary vcclor liad soybcc~~ t~-ypsin i~illib~to~ (.SljTi) as
tlie gcne of agrono~n~c ~ntercst, ~ipill as tllc re1ect;iblc marker arid ilitlA gcnc as reporter.
In both ~hc vectors ripill ;)lid iiidA genea ivc~c I't~hed ~nto ;I singlc unit tl~ougli their
products exli~bit indepe~ident activities. AM4 ehpla~its ivc~c co-cult~vatcd with
Agrobucicriirr~l stralli of lrltercst follow~rig tlie standard procedures. Ccfotaxiiiie was used
to terminate tlie growl1 of Agrobucieriu~~i and kanamyein was used for select~un of the
putative rransfornia~~ts. Starldtlrd~zed protocol of icgeneralio~l was used fu~ ~ege~ieration
and recovely of the tralfsgenlc plants. Hardelled and transplanted rra~isgcnics wcre